normfluodbf

Project status Science Liposomes Flash Liposomes Liposomes Demandez moi n'importe quoi ! Emojis

๐Ÿ‘จ๐Ÿพโ€๐Ÿ’ป ๐Ÿ“ฆ What is ?

The {normfluodbf} package is designed to clean and normalize DBF and DAT files obtained from liposome flux assay (LFA) experiments performed with the FLUOstar microplate reader. This package is designed to be multi-dimensional, meaning the expectation is that file types from experiments other than liposome flux assays should be compliant with this package if they meet the same requirements as the original files for which this package was designed.

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๐Ÿ’Š Package Context

What is the inspiration behind something like normfluodbf? First, the name of this package is misleading because it was initially designed to clean and normalize DBF files; however, the package is able to clean DAT files as well. The next sub-sections will shed more light on the context for the creation of this package. First, look at my abstract for the seminar I presented as a graduate student.

๐Ÿงฌ ๐Ÿ‘จโ€๐Ÿ”ฌ Abstract

Liposome flux assays (LFAs) provide a robust, affordable, and high throughput tool for the study of membrane channels. The project entailed adapting an LFA system from (Su et al., 2016) used in the study of Potassium (K) channels. The project conceptualized an LFA system centered around Voltage-gated sodium channels (NaVs), specifically the NavAb channel. NaVs are physiologically important in the generation of action potentials within systems that require excitability for function and are pathologically linked to conditions like epilepsy, migraines, etc. One reasoned that developing and testing an LFA system in the context of NavAb, if successful, will provide a good tool for screening molecules capable of NavAb interactions (in therapeutic or pathological scenarios). Interestingly, NavAb can conduct Na+ and H+ ions, amongst others, but the project one completed focused on the proton (H+) conductivity of the channel. One developed liposomes with NavAb insertions, and under specific conditions, I was able to detect proton (H+) conductivity through the channel driven by the K+ or Cs+ Nernst potentials. One utilized the fluorophore (ACMA) within the liposome as the proton flux signal indicator in all experimental conditions. It was confirmed that NavAb can conduct protons (H+), the NavAb LFA system worked, and it was found that increasing concentrations of ACMA (2-5 uM) within the acceptable range (0.2uM to 20uM) had no effect on proton quenching. An aberrant observation was that very high concentrations of ACMA (20uM) generated noise that was useful in determining noise-signal boundaries. One thought it was fascinating to find a system that could be understood or interpreted using noise as well as signal.

๐Ÿงช What are Liposome Flux Assays ?